Study of the effects of oral zinc supplementation on peroxynitrite levels, arginase activity and NO synthase activity in seminal plasma of Iraqi asthenospermic patients

Reproductive Biology and Endocrinology

Abdul Razzaq S Alsalman¹  Lamia A Almashhedy²  Mahmoud Hussein Hadwan² 

[1] Surgery Department, College of Medicine, Babylon University, Babylon, Iraq;Chemistry Department, College of Science, Babylon University, Babylon, Iraq

关键词: Oxidative stress;    Zinc supplementation;    Zinc;    Peroxynitrite;    Arginase;    Nitric oxide synthase;    Asthenospermia;   

Others  :  809714
DOI  :  10.1186/1477-7827-12-1

 received in 2013-11-21, accepted in 2013-12-29,  发布年份 2014

【 摘 要 】

Background

Low concentrations of nitric oxide (NO) are necessary for the biology and physiology of spermatozoa, but high levels of NO are toxic and have negative effects on sperm functions. Although several studies have considered the relationship between infertility and semen NO concentrations, no study on the effects of asthenospermia treatments such as oral zinc supplementation on concentrations of NO, which are important in fertility, has been reported. Studies have shown that oral zinc supplementation develops sperm count, motility and the physical characteristics of sperm in animals and in some groups of infertile men. The present study was conducted to study the effect of zinc supplementation on the quantitative and qualitative characteristics of semen, along with enzymes of the NO pathway in the seminal plasma of asthenospermic patients.

Methods

Semen samples were obtained from 60 fertile and 60 asthenozoospermic infertile men of matched age. The subfertile group was treated with zinc sulfate; each participant took two capsules (220 mg per capsule) per day for 3 months. Semen samples were obtained (before and after zinc sulfate supplementation). After liquefaction of the seminal fluid at room temperature, routine semen analyses were performed. The stable metabolites of NO (nitrite) in seminal plasma were measured by nitrophenol assay. Arginase activity and NO synthase activity were measured spectrophotometrically.

Results

Peroxynitrite levels, arginase activity, NO synthase activity and various sperm parameters were compared among fertile controls and infertile patients (before and after treatment with zinc sulfate). Peroxynitrite levels and NO synthase activity were significantly higher in the infertile patients compared to the fertile group. Conversely, arginase activity was significantly higher in the fertile group than the infertile patients. Peroxynitrite levels, arginase activity and NO synthase activity of the infertile patient were restored to normal values after treatment with zinc sulfate. Volume of semen, progressive sperm motility percentage and total normal sperm count were increased after zinc supplementation.

Conclusions

Treatment of asthenospermic patients with zinc supplementation leads to restored peroxynitrite levels, arginase activity and NO synthase activity to normal values and gives a statistically significant improvement of semen parameters compared with controls.

Trial registration

ClinicalTrials.gov identifier: NCT01684059

【 授权许可】

   
2014 Hadwan et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

【 参考文献 】

• [1]Brugh VM, Lipshultz LI: Male factor infertility: evaluation and management. Med Clin North Am 2004, 88:367-385.
• [2]Kamel RM: Management of the infertile couple: an evidence-based protocol. Reprod Biol Endocrinol 2010, 8:21. BioMed Central Full Text
• [3]Poland ML, Moghissi KS, Giblin PT, Ager JW, Olson JM: Variation of semen measures within normal men. Fertil Steril 1985, 44:396-400.
• [4]Guzick DS, Overstreet JW, Factor-Litvak P: Sperm morphology, motility, and concentration in fertile and infertile men. N Engl J Med 2001, 345:1388-1393.
• [5]Wong WY, Thomas CM, Merkus JM, Zielhuis GA, Steegers-Theunissen RP: Male factor sub-fertility: possible causes and the impact of nutritional factors. Fertil Steril 2000, 73:435-442.
• [6]Olayemi FO: A review on some causes of male infertility. Afr J Biotechnol 2010, 9(20):2834-2842.
• [7]Khan MS, Zaman S, Sajjad M, Shoaib M, Gilani G: Assessment of the level of trace element zinc in seminal plasma of males and evaluation of its role in male infertility. Int J App Basic Med Res 2011, 1:93-99.
• [8]Favier AE: The role of zinc in reproduction. Hormonal Mech. Biol Trace Elem Res 1992, 32:363-381.
• [9]Freedman LP: Anatomy of the steroid receptor zinc finger region. Endocr Rev 1992, 13:129-145.
• [10]Chimienti F, Aouffen M, Favier A, Seve M: Zinc homeostasis-regulating proteins: new drug targets for triggering cell fate. Curr Drug Targets 2003, 4:323-338.
• [11]Zago MP, Oteiza PI: The antioxidant properties of zinc: interactions with iron and antioxidants. Free Radic Biol Med 2001, 31:266-274.
• [12]Ozturk A, Baltaci AK, Bediz CS, Mogulkoc R, Gungor S: Effects of zinc and melatonin deficiency on testicular tissue of rats. Biol Trace Elem Res 2003, 96:255-262.
• [13]Lewis-Jones DI, Aird IA, Biljan MM, Kingsland CR: Effects of sperm activity on zinc and fructose concentrations in seminal plasma. Hum Reprod 1996, 11:2465-2467.
• [14]de Lamirande E, Gagnon C: Impact of reactive oxygen species on spermatozoa: a balancing act between beneficial and detrimental effects. Hum Reprod 1995, 10:15-21.
• [15]Sies H: Strategies of antioxidant defense. Eur J Biochem 1993, 215:213-222.
• [16]Sikka SC: Relative impact of oxidative stress on male reproductive function. Curr Med Chem 2001, 8:851-862.
• [17]Moncada S, Higgs A: The L-arginine-nitric oxide pathway. N Engl J Med 1993, 329:2002-2012.
• [18]Foghi K, Novin MG, Jabbari ZM, Najafi T, Heidari MH, Yasoori AR: Immuno-histochemical localization of endothelial nitric oxide synthase in testicular cells of men with nonobstructive azoospermia. Iran J of Reprod Med 2011, 9:277-280.
• [19]Carreira BP, Carvalho CM, Araújo IM: Regulation of injury-induced neurogenesis by nitric oxide. Stem Cells Int 2012, 2012(895659):15.
• [20]Wang Y, Goligorsky MS, Lin M, Wicox JN, Marsden PA: A novel, testis-specific mRNA transcript encoding an NH2-terminal truncated nitric oxide synthase. J Biol Chem 1997, 272:11392-11401.
• [21]Herrero MB, Gagnon C: Nitric oxide: a novel mediator of sperm function. J Androl 2001, 22:349-356.
• [22]Aquila S, Giordano F, Guido C, Rago V, Carpino A: Nitric oxide involvement in the acrosome reaction triggered by leptin in pig sperm. Reprod Biol Endocrinol 2011, 9:133. BioMed Central Full Text
• [23]Lee NPY, Cheng CYC: Nitric oxide and cyclic nucleotides. Their roles in junction dynamics and spermatogenesis. Oxid Med Cell Longev 2008, 1:25-32.
• [24]Amiri I, Sheikh N, Najafi R: Nitric oxide level in seminal plasma of fertile and infertile males and its correlation with sperm parameters. Daru 2006, 14:197-202.
• [25]Amiri I, Sheikh N, Najafi R: Nitric oxide level in seminal plasma and its relation with sperm DNA damages. Iran Biomed J 2007, 11:259-64.
• [26]Ghaffari MA, Rostami M: Lipid peroxidation and nitric oxide levels in male Smokers' spermatozoa and their relation with sperm motility. J Reprod Infertil 2012, 13:81-87.
• [27]Sheikh N, Amiri I, Najafi R, Goodarzi MT: The correlation between total antioxidant capacity and nitric oxide concentration in seminal plasma with sperm DNA damage. Afr J Biotechnol 2010, 9:5739-5745.
• [28]Lanzafame F: Oxidative stress and medical antioxidant treatment in male infertility. Reprod BioMed Online 2009, 19:638-659.
• [29]Ghasemi N, Babaei H, Azizallahi S, Kheradmand A: Effect of long-term administration of zinc after scrotal heating on mice spermatozoa and subsequent offspring quality. Andrologia 2009, 41:222-228.
• [30]Rafique M, Naqvi A, Nankani K: Zinc improves the quality of semen albino rats. Medical channel 2010, 16:619-622.
• [31]Wong WY, Merkus HM, Thomas CM, Menkveld R, Zielhuis GA, Steegers-Theunissen RP: Effects of folic acid and zinc sulfate on male factor subfertility: a double-blind, randomized, placebo-controlled trial. Fertil Steril 2002, 77:491-498.
• [32]Hadwan MH, Almashhedy LA, Alsalman AS: Oral zinc supplementation restore high molecular weight seminal zinc binding protein to normal value in Iraqi infertile men. BMC Urol 2012, 12:32. BioMed Central Full Text
• [33]Hadwan MH, Almashhedy LA, Alsalman AS: The key role of zinc in enhancement of total antioxidant levels in spermatozoa of patients with asthenozoospermia. Am J of Mol and Cell Biol 2013, 1:52-61.
• [34](WHO) World Health Organization: Laboratory Manual for the Examination of Human Semen and Semen-Cervical Mucus Interaction. 4th edition. Cambridge: Cambridge University Press UK; 1999:8-11.
• [35]Salter M, Knowles RG: Assay of NOS activity by the measurement of conversion of oxyhemoglobin to methemoglobin by NO. Nitric Oxide Protoc, Methods in Mol Biol, Vol 1998, 100:61-65. doi:10.1385/1-59259-749-1:61
• [36]Geyer JW, Dabich D: Rapid method for determination of arginase activity in tissue homogenates. Anal Biochem 1971, 39:412-417.
• [37]Beckman JS, Ischiropoulos H, Zhu L, van der Woerd M, Smith C, Chen J, Harrison J, Martin JC, Tsai M: Kinetics of superoxide dismutase and iron-catalyzed nitration of phenolics by peroxynitrite. Arch Biochem Biophys 1992, 298:438-445.
• [38]Szabo C: Multiple pathways of peroxynitrite cytotoxicity. Toxicol Lett 2003, 140–141:105-112.
• [39]Pryor WA, Jin X, Squadrito GL: One- and two-electron oxidations of methionine by peroxynitrite (hydroxyl radical/nitric oxide/ superoxide/ sfoide/ ethylene). Proc Natl Acad Sci USA 1994, 91:11173-11177.
• [40]Radi R, Beckman JS, Bush KM, Freeman BA: Peroxynitrite-mediated sulfhydryl oxidation: the cytotoxic potential of super-oxide and nitric oxide. J Biol Chem 1991, 266:4244-4250.
• [41]Castro L, Rodriguez M, Radi R: Aconitase is readily inactivated by peroxynitrite, but not by its precursor, nitric oxide. J Biol Chem 1994, 269:29409-29415.
• [42]Crow JP, Beckman JS, McCord JM: Sensitivity of the essential zinc-thiolate moiety of yeast alcohol dehydrogenase to hypochlorite and peroxynitrite. Biochemistry 1995, 34:3544-3552.
• [43]Loganathasamy K: Nitric oxide: a double edged weapon for sperm functions. J Vet Sci Technol 2012, 3:6.
• [44]Tsai HH, Hamilton TP, Tsai JHM, Beckman JS: Studies of peroxynitrite anion-water complexes. Struct Chem 1995, 6:323-332.
• [45]Ramya T, Misro MM, Sinha D, Nandan D, Mithal S: Altered levels of seminal nitric oxide, nitric oxide synthase, and enzymatic antioxidants and their association with sperm function in infertile subjects. Fertil Steril 2011, 95:135-140.
• [46]Abd-Elmoaty MA, Saleh R, Sharma R, Agarwal A: Increased levels of oxidants and reduced antioxidants in semen of infertile men with varicocele. Fertil Steril 2010, 94:1531-1534.
• [47]Köksal IT, Erdogru T, Gülkesen H, Sezer C, Usta M, Çiftçioglu A, Baykara M: The potential role of inducible nitric oxide synthase (iNOS) activity in the testicular dysfunction associated with varicocele: an experimental study. Int Urol Nephrol 2004, 36:67-72.
• [48]Chung HY, Seo KS, Rim JS: Effects of nitric oxide and peroxynitrite on sperm motility and viability. Korean J Urol 1998, 39:896-902.
• [49]Brown GC, Borutaite V: Nitric oxide inhibition of mitochondrial respiration and its role in cell death. Free Radic Biol Med 2002, 33:1440-1450.
• [50]Venkatakrishnan P, Nakayasu ES, Almeida IC, Miller RT: Arginase activity in mitochondria – an interfering factor in nitric oxide synthase activity assay. Biochem Biophys Res Commun 2010, 394:448-452.
• [51]Mori M, Gotoh T: Regulation of nitric oxide production by arginine metabolic enzymes. Biochem Biophys Res Commun 2000, 275:715-719.
• [52]Mori M: Regulation of nitric oxide synthesis and apoptosis by arginase and arginine recycling. J Nutr 2007, 137:1616S-1620S.
• [53]Lefevre PLC, Palin M-F, Murphy BD: Polyamines on the reproductive landscape. Endocr Rev 2011, 32:694-712.
• [54]Morales ME, Rico G, Bravo C, Tapia R, Alvares C, Mendez JD: Progressive motility increase caused by L-arginine and polyamines in sperm from patients with idiopathic and diabetic asthenozoospermia. Ginecol Obstet Mex 2003, 71:297-303.
• [55]Qiu CH, Ohe M, Matsuzaki S: Effect of diethylstilbestrol on polyamine metabolism in hamster epididymis. Asian J Androl 2003, 5:301-306.
• [56]Calandra RS, Rulli SB, Frungieri MB, Suescun MO, González-Calvar SI: Polyamines in the male reproductive system. Acta Physiol Pharmacol Ther Latinoam 1996, 46:209-222.
• [57]Srivastava S, Desai P, Coutinho E, Govil G: protective effect of L-arginine against lipid peroxidation in goat epididymal spermatozoa. Physiol Chem Phy Med NMR 2000, 32:127-135.
• [58]Belle NA, Dalmolin GD, Fononi G, Rubin MA, Rocha JB: Polyamine reduce lipid peroxidation induced by different pro-oxidant agents. Brain Res 2004, 1008:245-251.
• [59]Greene JM, Feugang JM, Pfeiffer KE, Stokes JV, Bowers SD, Ryan PL: L-arginine enhances cell proliferation and reduces apoptosis in human endometrial RL95-2 cells. Reprod Biol Endocrinol 2013, 11:15. BioMed Central Full Text
• [60]Méndez JD, Hernández MP: Effect of L-arginine and polyamines on sperm motility. Ginecol Obstet Mex 1993, 61:229-234.
• [61]Cai D, Deng K, Mellado W, Lee J, Ratan RR, Filbin MT: Arginase I and polyamines act downstream from cyclic AMP in overcoming inhibition of axonal growth MAG and myelin in vitro. Neuron 2002, 35:711-719.
• [62]Alsalman AR, Almashhedy LA, Hadwan MH: Zinc supplementation attenuates lipid peroxidation and increases antiperoxidant activity in seminal plasma of Iraqi asthenospermic men. Life Sci J 2013, 10:989-997.
• [63]Di Leo V, DInca R, Barollo M, Tropea A, Fries W, Mazzon E, Irato P, Cecchetto A, Sturniolo GC: Effect of zinc supplementation on trace elements and intestinal metallothionein concentrations in experimental colitis in the rat. DIG LIVER D 2001, 33:135-139.
• [64]Suriya J, Bharathiraja S, Sekar V, Rajasekaran R: Metallothionein induction and antioxidative responses in the estuarine poly chaeta Capitella capitata (Capitellidae). Asian Pacific J Trop Biomed 2012, 2:S1052-S1059.
• [65]Hrabak A, Bajor T, Temesi A: The inhibitory effect of nitrite, a stable product of nitric oxide (NO) formation, on arginase. FEBS Lett 1996, 390:203-206.
• [66]Reed DJ: Status of calcium and thiols in hepatocellular injury by oxidative stress. Semin Liver Dis 1990, 10:285-92.
• [67]Calabrese V, Cornelius C, Rizzarelli E, Owen J, Dinkova-Kostova A, Butterfield D: Nitric oxide in cell survival: a Janus molecule. Antioxid Redox Signal 2009, 11:2717-2739.